Asundi V, Cowan K, Matzura D, Wagner W, Dreher K L
Geisinger Clinic, Weis Center for Research, Danville, PA 17822.
Eur J Cell Biol. 1990 Jun;52(1):98-104.
Northern blot analysis using intraspecies and cross-species cDNA probes encoding a variety of proteoglycan (PG) core proteins was employed to examine in vitro and in vivo vascular smooth muscle cell (VSMC) extracellular matrix (ECM) PG gene expression. Similar studies were performed using a cDNA probe encoding the rat chondrosarcoma link protein. VSMCs maintained in vitro as well as in vivo expressed a 1.8 kb transcript homologous to a cDNA encoding the bovine bone small PGII. Densitometric analysis revealed an 11-fold increase in the in vivo level of this transcript when compared to an equivalent amount of RNA extracted from rat VSMCs maintained in vitro. Rat aorta medial tissue contained approximately a 10-fold higher level of the bovine bone PGII homologous transcript when compared to rat chondrosarcoma tissue. A 2.9 kb transcript homologous to a cDNA encoding the human bone small PGI (biglycan) was detected in poly(A)+ RNA isolated from rat VSMCs maintained in vitro as well as within rat thoracic aorta medial tissue. Densitometric analysis revealed a 0.55-fold increase in the in vitro level of the PGI transcript when compared to in vivo levels. Rat chondrosarcoma expressed approximately a 3-fold higher level of a human bone PGI homologous transcript when compared to rat aorta medial tissue. A 7.3 kb and an 8.4 kb transcript homologous to a cDNA encoding the rat chondrosarcoma large hyaluronic acid (HA) binding chondroitin-sulfate (CS) PG core protein was detected in poly(A)+ RNA extracted from rat VSMCs maintained in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)
利用编码多种蛋白聚糖(PG)核心蛋白的种内和种间cDNA探针进行Northern印迹分析,以检测体外和体内血管平滑肌细胞(VSMC)细胞外基质(ECM)PG基因的表达。使用编码大鼠软骨肉瘤连接蛋白的cDNA探针进行了类似研究。体外和体内培养的VSMC均表达与编码牛骨小PGII的cDNA同源的1.8 kb转录本。密度分析显示,与从体外培养的大鼠VSMC中提取的等量RNA相比,该转录本在体内的水平增加了11倍。与大鼠软骨肉瘤组织相比,大鼠主动脉中膜组织中牛骨PGII同源转录本的水平大约高10倍。在从体外培养的大鼠VSMC以及大鼠胸主动脉中膜组织中分离的聚腺苷酸加(poly(A)+)RNA中,检测到与编码人骨小PGI(双糖链蛋白聚糖)的cDNA同源的2.9 kb转录本。密度分析显示,与体内水平相比,PGI转录本在体外的水平增加了0.55倍。与大鼠主动脉中膜组织相比,大鼠软骨肉瘤中与人骨PGI同源转录本的表达水平大约高3倍。在从体外培养的大鼠VSMC中提取的poly(A)+ RNA中,检测到与编码大鼠软骨肉瘤大型透明质酸(HA)结合硫酸软骨素(CS)PG核心蛋白的cDNA同源的7.3 kb和8.4 kb转录本。(摘要截短于250字)